Mechanisms of Gene Regulation
zhao, hongwei , Li, Quinn .
A proteomic study of Arabidopsis cleavage and polyadenylation specificity factor.
In eukaryotic organisms, precursor messenger RNA (pre-mRNA) is processed to produce mature mRNA. The processing includes splicing, 5’ end capping, and polyadenylation. The process of polyadenylation involves addition of a polyadenosine tail to the 3’ end of an mRNA. The poly(A) tail protects the mRNA molecule from exonucleases degradation, and is required for exportation to cytoplasm and translation initiation. It has been demonstrated that the recognition of the polyadenylation signals, the cleavage, and the polyadenylation itself, need a group of essential proteins, named cleavage and polyadenylation specificity factor (CPSF).
In Arabidopsis thaliana, we have identified five CPSF genes that encode CPSF related proteins. Due to its essential role in RNA processing, CPSF has been hypothesized to work together with many other protein factors as a pre-mRNA processing complex, including polyadenylation, splicing, and potentially transcription termination factors. I am interested to test this hypothesis by isolating the protein complexes that are formed with CPSF subunits.
To achieve this goal, experiments designed to purify Arabidopsis CPSF and other closely interacting factors by tandem affinity purification method, followed by mass spectrometry identification, were carried out. Four known Arabidopsis CPSF factors were used as “baits” and four separate protein complexes were purified through their affinity interaction to those baits. Among those purified proteins, some are novel. Now the biochemical characterization of those protein factors is undergoing. The results will be presented and discussed.
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1 - miami university, botany department, oxford, ohio, 45056, usa
2 - miami university, botany department
tandem affinity purification
cleavage and polyadenylation specificity factor.
Presentation Type: Plant Biology Abstract
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM